PROJECT ABSTRACT Three million Americans have glaucoma with only half of them aware they carry the diagnosis. There is no cure and 10% of patients who with adequate treatment still have vision loss due to inadequate intraocular pressure control and compliance. Gene therapy as an intervention removes the responsibility of patient-driven treatments to address this global burden. The SIRT1 signaling pathway plays a role in visual preservation during optic nerve injury, making an excellent target gene to introduce this concept. This proposal requests support for a mentored career development award for an ophthalmologist with interest in developing cell- specific gene targeting using AAV to treat glaucoma. The expertise of the mentor involves the use of SIRT1 in a glaucoma model based on evidence from other models of optic nerve injury. At the completion of the award period the candidate will gain valuable experience required to propose, construct, and execute experiments in visual research from patient individualized data gained from current studies at UPenn. The candidate?s goal is to lead a research program that addresses a therapeutic need in glaucoma through the use of gene therapy. This skill set offers a novel and rational approach to treat other cells in eye to manage glaucoma in the future. This project is under the supervision of senior investigators at UPenn in the departments of Ophthalmology, Physiology, and the Center for Advanced Retinal and Ocular Therapeutics (CAROT) who will provide resources for candidate?s career development. The plan involves regular scheduled meetings, structured coursework, guidance for support, and innovative and translational scientific aims. All of the mentors have secured funding to provide necessary resources to ensure success of the aims in the proposal. The institution, mentors, and collaborators outlined in this proposal are dedicated to the research and growth of early investigators. This plan will position the candidate for independence and specialized expertise at the end of the award period through execution of the following AIMS: AIM 1) Define the molecular mechanism of SIRT1 pathway on RGCs. We will confirm the role of SIRT1 pathway expression in induced pluripotent stem cell (IPS) derived RGCs from unaffected control patients using cell stress from hydrogen peroxide (H2O2) and a stretch chamber. AIM 2) Determine the role of SIRT1 signaling with small molecular activators and targeted AAV expression in a mouse model of glaucoma. We will evaluate the role of SIRT1 activation using a mouse model of chronic glaucoma. AIM 3) Develop cell specific adenoviral specific gene delivery system for microglia and astrocytes. We will develop the technology to deliver SIRT1 target gene to astrocytes and microglial cells with specific cell promotors using in vivo models of chronic glaucoma.